VALIDATION IMAGES
Formalin-fixed and paraffin embedded human breast carcinoma labeled with Anti-phospho-eIF4EBP1(Thr37/46) Polyclonal Antibody, Unconjugated (bs-3019R) followed by conjugation to the secondary antibody and DAB staining
Paraformaldehyde-fixed, paraffin embedded mouse small intestine tissue; Antigen retrieval by boiling in sodium citrate buffer(pH6) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (normal goat serum) at 37°C for 20min; Antibody incubation with Rabbit Anti-eIF4EBP1 Polyclonal Antibody, Unconjugated (bs-3019R) at 1:400 overnight at 4°C, followed by a conjugated secondary and DAB staining
Image on the left is the same conditions, with the peptide incubated with the antibody prior to staining.
Mouse ovary and Rat ovary lysates probed with eIF4EBP1 (Thr37+Thr46) Polyclonal Antibody, unconjugated (bs-3019R) at 1:300 overnight at 4°C followed by a conjugated secondary antibody at 1:10000 for 90 minutes at 37°C.
HepG2 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with phospho-eIF4EBP1 (Thr37 + Thr46) Antibody(bs-3019R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
HepG2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-eIF4EBP1 (Thr37 + Thr46)) polyclonal Antibody, Unconjugated (bs-3019R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (phospho-eIF4EBP1 (Thr37 + Thr46)) polyclonal Antibody, Unconjugated (bs-3019R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.